环境激素烯菌酮引发小鼠前列腺癌细胞的毒性机制

为探讨环境激素烯菌酮引发小鼠前列腺癌(RM-1)细胞毒性机制,设计了3组实验,分别为对照组、烯菌酮染毒组(染毒组)和烯菌酮染毒后添加维生素E组(实验组),各组给药培养48h后,观察细胞形态变化。检测细胞增殖活性、丙二醛(MDA)含量、抑制羟自由基能力、乳酸脱氢酶(LDH)活性、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性。结果表明,与对照组相比,染毒组RM-1细胞圆缩脱落明显,贴壁细胞减少;细胞增殖率、抑制羟自由基能力、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性明显降低(p<0.05),LDH活性和MDA含量显著升高(p<0.05)。与染毒组相比,染毒后添加维生素E,圆缩脱落细胞减少,而梭形贴壁细胞明显增加;细胞增殖率、抑制羟自由基能力、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性显著升高(p<0.05),LDH活性和MDA含量明显下降(p<0.05)。这表明维生素E可能通过其抗氧化性拮抗了烯菌酮引发的氧化应激,缓解了细胞膜的损伤,维持了细胞膜的完整性和膜蛋白的功能,防止了细胞内LDH外逸、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的下降,从而逆转了烯菌酮引发的细胞死亡和增殖率的下降。     

Observations on the reproductive cycles of Monodonta lineata,Gibbula umbilicalis and G. cineraria

Monthly samples of gonads of 3 common species of intertidal trochids, Monodonta lineata (da Costa), Gibbula umbilicalis (da Costa) and G. cineraria (L.) were fixed for examination by light microscopy. Numbers and proportions of immature and mature oocytes in sections of ovaries were determined. In M. lineata and G. umbilicalis, the annual reproductive cycles were very similar, the testis and ovary of both species being empty brown structures from September to January/February, when gametogenesis began and the gonads began to swell and turn green in both sexes. As ripening continued, the ovary remained green in G. umbilicalis, but in M. lineata turned greygreen by the end of May. The testis of both species changed, to yellowish in G. umbilicalis and creamy-pink in M. lineata. Spawning occurred throughout summer and early autumn, and the gonads reverted to their former empty state. These changes were matched by the increase and subsequent decrease in proportion of ripe oocytes in the gonad. Some ripe oocytes are not shed during the spawning season, but no evidence of cytolysis of such oocytes was found. In G. cineraria, in contrast, the ovary remained green and swollen and the testis remained creamy-white and packed with sperm throughout the year. The proportion of ripe to unripe oocytes in the ovary fluctuated for the entire sampling period, and did not indicate the time of spawning. In the testis of all three species, spermatogonia were rarely seen, and spermatid stages were uncommon.     

Responses of ovaries and testes of Lytechinus variegatus (Echinodermata: Echinoidea) to dietary administration of estradiol, progesterone and testosterone

 In many vertebrates, environmental factors influence gamete differentiation and growth of the mature gonad through alteration of sex steroid production or action; however, it is unclear how gamete differentiation and gonadal growth are regulated in echinoids. The purpose of this study was to examine the influence of dietary administration of estradiol (E2), progesterone (P4), testosterone (T) and finasteride (F, a 5α-reductase inhibitor) on the ovaries and testes of mature Lytechinus variegatus (Lamarck) during gonadal growth. Echinoids were fed a formulated diet supplemented with steroids or steroids in combination with finasteride for 36 d. The effects of dietary administration of steroids on L. variegatus were both steroid- and sex-specific. The mean ovary index was 54% greater in individuals fed E2 than from individuals fed the control (C) diet (10.0 ± 1.1 vs 6.5 ± 0.7, respectively; P < 0.05). Individuals fed E2, P4, E2/P4, and P4 in combination with F had significantly smaller oocytes (P < 0.005) than individuals fed C. The volume fraction occupied by nutritive phagocytes in ovarian tubules from individuals fed E2/P4 or P4 were significantly larger than the volume fraction occupied by nutritive phagocytes from individuals fed C, indicating that E2 and 5α-reduced progestins may promote nutrient allocation to nutritive phagocytes. Although oocytes from the individuals fed T alone were significantly smaller than those fed T in combination with F, oocytes from individuals in both treatments were significantly larger compared to oocytes from individuals fed C. These data suggest that upon removal of 5α-reduced androgens, T is able to promote an increase in oocyte diameters. In contrast, the mean testis index was 56% greater in individuals fed P4 than in individuals fed C (8.9 ± 0.6 vs 5.7 ± 0.9, respectively; P < 0.05); the testis index did not increase in individuals fed P4/F, suggesting that 5α-reduced progestins stimulate testicular growth. Testes growth in the presence of 5α-reduced progestins was accomplished by significant increases in the volume fraction occupied by nutritive phagocytes and by a significant reduction in the volume fraction occupied by spermatogenic columns in testicular tubules. These data further indicate that 5α-reduced progestins (or 5α-reduced androgens) may inhibit spermatogenic column formation. In conclusion, E2 stimulated ovarian growth but inhibited oocyte growth, whereas T had no affect on ovarian growth but promoted oocyte growth in L. variegatus. We hypothesize that the E2 (or E2 metabolites) and/or 5α-reduced androgens in combination with T regulate oocyte growth in the echinoid L. variegatus. In addition, 5α-reduced progestins promoted nutrient accumulation in nutritive phagocytes within the ovaries and the testes. Furthermore, 5α-reduced progestins stimulated growth of the testes and inhibited spermatogenic column formation, suggesting that 5α-reduced progestins regulate nutrient accumulation into nutritive phagocytes and spermatogenic column formation in L. variegatus. The differences in estrogen effects between echinoids and asteroids may be related to differences in gonad morphology and, ultimately, the differences in cellular signalling pathways (paracrine vs endocrine). Received: 22 May 1999 / Accepted: 24 May 2000     

Evaluation of antioxidant potentials of Morinda morindoides leaf extract

This study investigated antioxidant status of animals given aqueous extract of Morinda morindoides leaves using the levels of reduced glutathione, total-thiol, vitamin C, and vitamin E as well as malondialdehyde concentrations as indices, and its in vitro antioxidant capacity. Thirty rats divided into five groups were used. Group A served as control and were administered distilled water while groups B, C, D, and E were given 100, 200, 400, and 800 mg per kilogram body weight of water-extracted constituents of M. morindoides for 28 days. Total phenolic compounds amounted to 83.6 ± 5.9 mg g^?1 gallic acid equivalent, while total flavonoid content was 9.5 ± 0.9 mg g^?1 pyrocathecol equivalent. Malondialdehyde in plasma was significantly decreased in a dose-dependent manner, ranging from 21% in groups B and C to 84% in groups D and E. Vitamins C and E were significantly increased, in group E by 91% and 17% compared with control. Total thiols and glutathione in plasma were significantly increased, with group E having 2.5-fold and 4.2-fold higher values than control.     

Vitamin E supplementation enhances growth and condition of nestling barn swallows (Hirundo rustica)

To optimize fitness, organisms may have to trade the number and quality of individual offspring against their own condition and survival. Limiting micronutrients such as antioxidants may be crucial to this trade-off. We investigated whether vitamin E, a major antioxidant in the diet of vertebrates, is limiting to barn swallow (Hirundo rustica) nestlings. We manipulated brood size to alter the intensity of sib–sib competition and supplemented nestlings with two different physiological doses of vitamin E while establishing a control group. Treatment effects were measured on body mass and size, feather growth, T cell-mediated immune response and hematocrit. Supplementation with vitamin E at intermediate physiological doses improved nestling mass and condition and feather growth, whereas higher physiological doses did not enhance offspring quality compared to a control treatment. The positive effects of vitamin E on body mass and condition were only detectable from days 6 to 10 when maximum growth rate is attained. Experimental enlargement of broods reduced body mass and size and T cell-mediated immune response only during the late nestling period. The effect of vitamin E supplementation did not depend on brood size manipulation, as revealed by the nonsignificant statistical interaction. This result contradicts the hypothesis that availability of vitamin E depends on intrabrood competition and instead suggests that it depends on concentration of vitamin E in the insect prey of swallows. Thus, antioxidants may be available in limited amounts to barn swallow nestlings and such limitation affects growth. In addition, present results confirm that barn swallow parents trade progeny number against growth and immunity of individual offspring.     

Evaluation of vitamin E and calcium effects on fluoride toxicity-induced fertility impairment

Chronic fluoride (Fl) toxicity is a serious public health problem globally where drinking water contains more than 1 ppm of Fl. Sodium fluoride (NaF) produced male reproductive system toxicity. The aim of the present study was to evaluate the amelioration of Fl toxicity-induced fertility impairment by vitamin E and calcium during the withdrawal period. The study was carried out on 70 adult male albino rats divided into five main groups: group I control; subdivided into group Ia (maintained on standard diet and water ad libitum for 60 days) and group Ib (maintained on standard diet and water ad libitum for 120 days), group II was administered NaF and subdivided into group IIa (administered NaF for 60 day and sacrificed) and group IIb (administered NaF for 60 day then maintained on standard diet and water ad libitum for a further 60 days), and treated groups III, IV, and V were administered NaF. Rats were maintained during withdrawal from NaF, on vitamin E (10 mg kg^?1 day^?1 for 60 days), calcium (50 mg kg^?1 day^?1 orally for 60 days), and both vitamin E and calcium, respectively. The duration of NaF administration was 60 days at a dose 20 mg kg^?1 day^?1 for all treated groups. The following parameters were determined: body and organ weights, sperm motility, sperm morphology, sperm viability, fertility test, and hormone assays: testosterone, in vitro testosterone production, luteinizing hormone, and follicular stimulating hormone. The combined administration of vitamin E and calcium during withdrawal from NaF showed significant improvement from chronic FL-induced toxicity on male reproductive organs.     

Ovarian morphology and oogenesis in Aurelia aurita (Scyphozoa: Semaeostomae): ultrastructural evidence of heterosynthetic yolk formation in a primitive metazoan

In Aurelia aurita, the ovaries arise as horseshoe-shaped evaginations of the gastrodermis in the floor of four interradial gastric pouches. Germ-cell islands arise within endodermally-derived gastrodermal cells. Oocytes grow and gradually bulge into the mesoglea while maintaining physical contact throughout vitellogenesis with specialized cells called trophocytes. Ultrastructural changes suggest that these cells transport yolk precursors from the coelenteron to the oocytes in a manner similar to that reported for the trophonema cells of anthozoan ovaries. Vitellogenesis involves both the autosynthetic activity of the oocyte organelles (Golgi complex and rough endoplasmic reticulum) and the heterosynthetic incorporation of precursors through endocytotic processes involving both coated pits and vesicles and smooth-surfaced tubules. Ultrastructural data suggest that different types or classes of yolk precursors enter the oocyte through the trophocytes and via the surrounding mesoglea. To our knowledge, this is the most primitive animal in which this type of yolk synthesis has been described. The trophocyte-oocyte relationship in oocytes of A. aurita is reminiscent of the trophonema-oocyte relationship in anthozoans and supports the belief that the Anthozoa and Scyphozoa share a close phylogenetic relationship.     

Ultrastructure of the ovary and oogenesis in the jellyfish Linuche unguiculata and Stomolophus meleagris,with a review of ovarian structure in the Scyphozoa

Ovarian structure and oogenesis has been examined in six scyphozoan species including the semaeostome Diplumularis antarctica Maas, 1908 (collected in 1987 in McMurdo Sound, Antarctic), the rhizostomes Cassiopea xamachana Bigelow, 1892 (collected in Belize in 1988), and Stomolophus meleagris L. Agassiz, 1862 (collected in Ft. Pierce Inlet in 1988), and the coronates Periphylla periphylla (Peron and Lesueur, 1810), Nausithoe atlantica Broch, 1914 (both collected in the Bahamas in 1988), and Linuche unguiculata (Schwartz, 1788) (collected in Nassau Harbor, Bahama Islands in 1989). Based on these findings and information on five other scyphozoan species from additional literature sources, at least two fundamentally different types of ovaries exist in the Scyphozoa. In semaeosotome and rhizostome species, oocytes develop in close association with specialized gastrodermal cells called trophocytes which may serve a nutritive function. However, coronate species lack trophocytes and oocytes develop freely in the mesoglea. The ovaries of S. meleagris and L. unguiculata are used as models to represent the ultrastructural events occurring during oogenesis in species having trophocytes and those lacking them, respectively. In both L. unguiculata and S. meleagris, the ovaries arise as evaginations of the gastrodermis in the floor of interradial pouches. Germ cells appear to originate from endodermally-derived gastrodermal cells and migrate into the mesoglea prior to vitellogenesis. In L. unguiculata, the oocytes develop freely within the mesoglea throughout vitellogenesis, while in S. meleagris each oocyte maintains contact with specialized gastrodermal cells called trophocytes. In the vitellogenic oocytes of both species, numerous invaginations of the oolemma result in the formation of intraooplasmic channels throughout the ooplasm. These channels are intimately associated with cisternae of rough endoplasmic reticulum and may play some role in yolk precursor uptake by substantially increasing the surface area of the oocyte. Vitellogenesis is similar in both species and involves the autosynthetic activity of the Golge complex and rough endoplasmic reticulum, and the heterosynthetic incorporation of yolk precursors through receptor-mediated endocytosis. However, in the oocytes of S. meleagris, the trophocytes probably play a role in the transfer of nutrients from the gastrovascular cavity to the oocyte. The present study suggests that scyphozoans were among the first metazoans to develop ovarian accessory cells during their reproductive evolution. The trophocyte-oocyte association observed in some scyphozoans is similar to but structurally less complex than the trophonema-oocyte association described from anthozoans. Scyphozoan ovarian morphology helps support the view that the Scyphozoa share a closer phylogenetic relationship with the Anthozoa than with the Hydrozoa.     

Amelioration of fluoride toxicity by vitamins and calcium on reproductive organs of female rat

Normal female rats of Wistar strain (Rattus norvegicus) weighing between 150–200 g were treated with fluoride (Fl) contaminated drinking water (FW, 5.8 ppm), vitamin C (6 mg) and vitamin C (6 mg) + D (6 mg once a week) + calcium (6 mg) for 30 days. Fl water treatment to rats produced reduction in weights of ovaries, uterus, vagina, kidneys, and adrenal glands, circulating levels of estrogen, number of litters, fertility rate, and altered tissue and serum biochemistry compared to control rats. However, cholesterol concentrations of ovaries and adrenals increased significantly. The above altered parameters were restored partially/completely after exogenous feeding with vitamin C and vitamins (C + D) and calcium. The data suggest that Fl-induced adverse effects on reproductive and other organs in female rats, whereas vitamin C, vitamin D and calcium treatment ameliorated Fl toxicity. Therefore, vitamins (C and D) and calcium play an important role in prophylactic treatment of fluorosis.     

Spawning pattern and type of fecundity in relation to ovarian allometry in the round herring <Emphasis Type="Italic">Etrumeus teres</Emphasis>

Spawning pattern (assessed by seasonal changes in ovarian developmental stages) and type of fecundity (assessed by analysis of oocyte-size frequency distributions) of the round herring Etrumeus teres were studied in relation to ovarian growth and seasonal changes in the gonadosomatic (GSI), hepatosomatic (HSI) and liposomatic (LSI) index as well as the somatic condition of spawners (CS) in a spawning ground of southern Japan. Except for summer, mature and recently spawned ovaries occurred all year round. Oogonia and primary oocytes were present in all ovaries, and cortical alveoli stage (CA) oocytes occurred in all mature, hydrated and partially spent (PS) females (PS: females containing post-ovulatory follicles). Before hydration, a clutch of larger yolked oocytes, undergoing synchronous growth (range 0.7–1.1 mm), was present in mature ovaries which was completely separated from a more heterogeneous clutch of oogonia, primary and secondary oocytes (<0.150 mm) and oocytes in the CA stage (range 0.15–0.60 mm). As vitellogenesis progressed, the yolked clutch increased in size but the CA oocytes remained arrested. The latter entered into the secondary growth phase when hydration started in the advanced batch. Ovarian growth was isometric in all developmental stages, validating the use of GSI, which showed a consistent monthly evolution among years. Spawning stopped in summer (July and August) and peaked in winter and spring. HSI correlated positively with GSI on both a monthly mean basis (r = 0.76) and individual fish basis (liver weight explained 67–83% of the variability in ovary weight when females were grouped into 1-unit GSI intervals) suggesting a significant role of liver in vitellogenesis. LSI and CS also showed marked seasonal changes peaking from summer to middle autumn. Overall results suggest that E. teres is a multiple spawner with a group-synchronous ovarian development and indeterminate annual fecundity, with the three processes linked to an isometric growth of the ovary. We propose that such a reproductive pattern is an adaptation to produce batches of large pelagic eggs through a protracted spawning season.     

Potential hepatoprotective effects of vitamin E and Nigella sativa oil on hepatotoxicity induced by chronic exposure to malathion in human and male albino rats

Malathion is an organophosphorus (OP) insecticide and has a wide range of use in agriculture, veterinary medicine, and public health. Malathion and other OP insecticides produce hepatotoxic effects. The objective of the present study was to investigate the protective effects of Nigella sativa oil and α-tocopherol (vitamin E) on the hepatotoxicity induced by malathion on workers involved in the formulation of pesticides, chronically exposed to malathion, and in male albino rats orally administrated malathion. This study was conducted on both human and experimental animals, the human study was conducted on 30 control subjects working as administrators and 45 subjects working in formulation of pesticides and exposed to malathion (≥3 years), all were males with age ranges from 30 to 60 years. The 45 males working in pesticides formulation were classified into three groups; (1) 15 workers exposed to pesticides (2) 15 workers exposed to pesticides and received vitamin (E), in a dose of 10 mg kg?1 day^?1 orally for 60 days, and (3) 15 workers exposed to pesticides and received 100 mg kg^?1 day^?1 of N. sativa oil for 60 days. The animal experiment was conducted on 40 adult male albino rats weighing 150–200 g. They were divided into four groups (10 rats in each group). First group served as the control group, the second group received malathion in a dose of 50 mg kg^?1 orally per day for 60 days, the third group received malathion (in the same dose and route of administration) and vitamin E in a dose of 10 mg kg^?1 day^?1 orally for 60 days, and the fourth group received malathion (in the same dose and route of administration) and N. sativa oil in a dose of 100 mg kg^?1 day^?1 orally for 60 days. Liver function tests (alanine aminotransferase [ALT], aspartate aminotransferase [AST], serum alkaline phosphatase [ALP], albumin, globulin, albumin/globulin ratio, and total proteins), antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD), and total glutathione peroxidase (GPx)), and lipid peroxidation [MDA] were analyzed in both human and animal experiments. The results of both human and animal study revealed that, exposure to malathion produced significant increases in AST, ALT, and lipid peroxidation. There were significant decrease in albumin, albumin/globulin ratio, total protein, and antioxidant enzymes. There was no significant change in ALP. In addition exposed workers showed significant decreases in serum globulin. Nigella sativa oil or vitamin E administration showed significant improvement of liver function tests, lipid peroxidation, and antioxidant enzymes impairment induced by malathion. Thus, dietary supplement, N. sativa oil, or vitamin E may represent a potential therapeutic agent in reducing malathion-induced hepatotoxicity.     

原花青素对镉引起的鸡胚睾丸氧化损伤的缓解作用

为探索原花青素对镉引起的鸡胚睾丸氧化损伤的缓解作用,本实验设置了对照组、镉、葡萄籽提取物原花青素(grape seed proanthocyanidin extract,GSPE)和镉+GSPE组,在胚胎期第6.5天(E6.5)注射0.05 mg·kg-1镉或2.5 mg·kg-1GSPE,统计孵化率和睾丸指数,并在E17.5检测睾丸形态、氧化指标、细胞凋亡和内质网应激相关基因的表达情况。结果表明:与对照组相比,镉处理组孵化率和睾丸指数均显著降低;睾丸组织结构出现了细胞核固缩、空泡化和染色质周缘化等变化,过氧化氢和丙二醛水平升高、超氧化物歧化酶活性显著降低、谷胱甘肽含量明显减少,凋亡细胞显著增多;睾丸中抗凋亡基因Bcl-2 mRNA表达水平下降,同时促凋亡基因Caspase3和内质网应激相关基因(XBP-1和HO-1)mRNA表达水平显著上升。然而,GSPE联合处理,明显缓解了镉引起的睾丸损伤,使孵化率和睾丸指数回升,抗氧化水平和内质网应激得到改善,凋亡细胞显著减少,睾丸形态趋于正常。结果表明:GSPE可通过其抗氧化作用降低XBP-1相关的内质网应激基因的表达,从而缓解镉引起的鸡胚睾丸生殖毒性。     

Annual reproductive cycles of the commercial sea urchinParacentrotus lividus from an exposed intertidal and a sheltered subtidal habitat on the west coast of Ireland

Reproduction of the commercial sea urchinParacentrotus lividus (Lamarck) from contrasting habitats on the west coast of Ireland was examined from May 1986 through August 1988. Urchins were collected intertidally from an exposed rocky shore and subtidally from a protected bay. Monthly measurements of the gonad index and histological examination of the gonads demonstrated thatP. lividus has an annual reproductive cycle. Although the two populations exhibited similar reproductive patterns over three breeding seasons, there were inter-annual and inter-population differences in the amplitude of gonadal growth and in the time when spawning started. The subtidal urchins had significantly larger gonads and exhibited a longer period of reproductive maturity compared with the intertidal urchins. This difference was also evident in the histology of the ovaries at the beginning of breeding, when most of the subtidal females contained mature ovaries, whereas most of the intertidal females contained premature ovaries. An inter-annual difference in the onset of spawning was observed, with the start of gamete release differing by as much as four weeks between years. It appears that inter-annual differences in sea temperature influence the temporal variation in spawning byP. lividus and that increasing temperature may serve as an exogenous cue for gamete release. The inter-annual variability in the onset of spawning suggests that photoperiod does not cue gamete release. Gonadal growth, on the other hand, occurs during the coldest months of the year and during the period of shortest days, suggesting that temperature and photoperiod may both influence gonadal growth during the winter. Oogenesis and spermatogenesis ofP. lividus were examined histologically. During the post-spawning recovery and growth stages, the gonads gained weight through growth of the nutritive phagocytes and the accumulation of periodic acid Schiff (PAS) + droplets by these cells. The PAS + material appears to play a nutritive role in gametogenesis. For the females, the frequencies of six ovarian maturity stages was assessed at approximately monthly intervals. Small oocytes were present throughout the year and clusters of early oocytes were most apparent during the spent and recovery stages. With the onset of vitellogenesis and subsequent accumulation of ova, the nutritive phagocytes and their PAS + droplets became depleted. The ovarian condition at the onset of breeding was variable, due to differences in the number of vitellogenic oocytes, differences in the number of ova in storage, and differences in the amount of PAS + material. In general, the nutritive phagocyte tissue is reduced by the onset of spawning and is exhausted by the termination of breeding. A similar series of events occurs during spermatogenesis. The relevance of this study for the use ofP. lividus as a brood-stock organism for mariculture is discussed.     

First ovarian maturity and recovery response in common carp, Cyprinus carpio after exposure to carbofuran

The present study embodies the observations on duration (45, 75 and 105 days) dependent effects of toxicologically low concentration (16 ppm) of a carbamate pesticide, carbofuran, on first ovarian maturity with correlative recovery response by withdrawal of treatment for 30 days in virgin yearlings of common carp, Cyprinus carpio var. Communis. In control fish histological organization of ovary and changes in gonado-somatic index (GSI) revealed the initiation of oocyte differentiation and development at the commencement of experiment in December-January and peak of maturation with fully grown yolky oocytes was attained by the time of termination of experiment in the end of March. In carbofuran exposed groups ovaries exhibited retardation of development and growth evident by significantly decreased GSI and predominace of immature oocytes passing through nuclear-cytoplasmic degeneration of variable magnitude. Severity of degenerative changes was more pronounced with prolonged duration. In recovery group ovaries had experienced regeneration as clued by the presence of oocytes of differential stages including vitellogenic and fully grown oocytes. In liver histopathological changes coupled with significant decrease in hepato-somatic index (HSI) attributed hepatic inactivation in all exposure groups whereas in recovery group an increase in HSI value with marked sign of hepatic reactivation was evidenced. No significant change in values of condition (CF) and somatic condition (SCF) factors substantiated the over all normal health of fish. Lipid and water content of ovary, liver and muscle registered the significant decrease at 105th day of exposure with concomitant augmented value for recovery group. These observations suggest that the commercial formulation of carbofuran, even at this toxicologically low concentration, is equally capable of affecting the first ovarian maturity with associated physiological implications but with reversibility in toxicity by withdrawal of pesticide exposure attributing regeneration in affected organs in due course time.     

Gonadal development and infrequent sex change in a population of the humbug damselfish,Dascyllus aruanus,in continuous coral-cover habitat

The expression of protogyny often differs among populations of a single species. The humbug damselfish, Dascyllus aruanus (Linnaeus), typically lives in spatially discrete groups of several individuals around live coral colonies and has been reported to be protogynous. The size and sexual composition of groups of this species, however, vary with the size of the coral patch or the degree of coral cover. The present study examined gonadal development and sexual pattern in a population of D. aruanus living in continuous coral-cover habitat. Fish were collected in Tumon Bay, Guam (13°31′N; 144°47′E), during a 2-week period in July and August 1996. Reflecting their distributions, small fish (n=54) were collected at depths of 1–2 m over continuous Porites spp. patches, and large fish (n=56) were collected at depths of 2–3 m over a continuous Acropora spp. patch. Gonadal development was examined histologically. Gonads of D. aruanus first developed an ovarian lumen and primary-growth stage oocytes. From this ovarian state or from more developed ovaries with cortical-alveolus stage oocytes, some gonads developed into testes through degeneration of oocytes and development of spermatogenic tissue. Developing spermatogenic tissue only occurred in gonads having pre-cortical alveolus stage oocytes, and the size of individuals with gonads of this type overlapped mainly with the size range of individuals with gonads containing only pre-vitellogenic oocytes (i.e. immature females). This suggested that functional female-to-male sex change was rare or absent in this population of D. aruanus. Moreover, the different habitat distributions of small and large fish suggest that sex determination in immature fish occurs in the absence of adults.     

Testicular damage after chronic exposure to carbendazim in male goats

The present investigation was aimed to determine the effect of chronic exposure of carbendazim on cellular changes in testes of male goats. The goats were randomly divided into two groups, control and treatment (N?=?7 each). The treatment group was exposed to carbendazim at the dose rate of 50?mg?kg^?1 body weight per day orally, once daily for 180 consecutive days. Testes were removed from control and experimental animals surgically on the 90th, 120th, and 180th day. On the 180th day of the treatment, a maximum number of seminiferous tubules became atrophic, and vacuolization of germinal epithelium, and sloughing of the germinal epithelium and sertoli cells was more marked as compared to days 90 and 120. Some of the seminiferous tubules were devoid of germinal cells. These pathological findings were supported by ultrathin and electron microscopic examination. On the basis of the present investigation it could be concluded that chronic exposure of carbendazim in male goats did not allow proper maturation of testis.     

Relationships between the movements,growth, age structures,and reproductive biology of the teleosts Sillago burrus and S. vittata in temperate marine waters

Sillago burrus and S. vittata both use sheltered, nearshore shallow waters ( 1.5 m) as nursery areas. However, the juveniles of the former species remain there for only a few months, before migrating into deeper waters (5 to 15 m) as they increase in size, whereas some juvenile S. vittata do not undergo a similar migration until considerably later. S. burrus rarely exceeded 2 yr of age and was never found beyond 4 yr of age. Although only a small number of S. vittata exceeded 2 yr of age, a few individuals of this species were caught between 4 and 7 yr old. The maximum and asymptotic lengths of S. burrus (251 and 180 mm, respectively) were far lower than those of S. vittata (325 and 320 mm, respectively), whereas the growth coefficients (K) were much higher for the former species, i.e. 2.4 vs 0.4. Virtually all S. burrus, and also those S. vittata that moved into deeper waters early in life, spawned at the end of their first year of life. Since relatively few S. burrus reached 2 yr of age, the attainment of almost full size by the end of their first year of life enables a relatively large number of eggs to be produced by fish at the end of their first year-for many, their only spawning period. Those individuals of S. vittata that remained in their shallow nursery areas until the end of their first year of life, did not reach maturity until the end of their second year of life. The proportions of mature gonads and the numbers of yolk-vesicle and yolk-granule oocytes and post-ovulatory follicles in ovaries were far higher in both S. burrus and S. vittata during December to February than in any other month, demonstrating that these two species spawn largely in these summer months. During this period, the ovaries of individual S. burrus and S. vittata often contained post-ovulatory follicles, as well as yolkvesicle and yolk-granule oocytes that ranged widely in size, strongly suggesting that both species are multiple spawners. During the spawning period of S. burrus, the ovaries possessed large numbers of hydrated oocytes and no post-ovulatory follicles or the reverse situation, and the oocytes tended to form several relatively discrete size groups. This indicates that S. burrus produces eggs in batches and that the spawning of the members of this species is synchronised. The presence of large numbers of yolk-granule oocytes with migrating nuclei in the ovaries of many S. vittata at certain times suggests that this species is also a batch- and synchronised spawner. Comparisons between the results of the present study and past work emphasise that the relationships between the timing of offshore movements and the sizes and ages at first maturity vary considerably amongst whiting species.     

Obligate parthenogenesis and reproductive division of labor in the Japanese queenless ant Pristomyrmex pungens

Summary Two types of workers were recognized in colonies of Pristomyrmex pungens: extranidal workers (which characteristically walk outside the nest) and intranidal workers (which characteristically stay inside the nest). The ovaries of extranidal workers showed little activity, whereas those of intranidal workers showed high activity and often contained mature oocytes. I therefore conclude that only the intranidal workers reproduce. A behavioral repertoire of 103 individuals was obtained and used to infer group subdivision using cluster analysis; in addition, principal component analysis was performed on the intranidal workers in this set. These data enabled objective separation of extranidal and intranidal workers. Intranidal workers were larger in size on average than extranidal workers; however, the distributions overlapped. Three tests for further subdivision within the group of intranidal workers indicated that such subdivision is weak, and it is also likely that all intranidal workers lay eggs. There was no significant correlation between body size and reproductive status. The number of mature oocytes per ant fitted a Poisson distribution, and the first two principal component factors scores of behavior showed significant correlation with head width. All extranidal workers had resorbed ovaries and also had yellow bodies (which indicated a history of oviposition). When and how the differentiation between the reproductive intranidal workers and the non-reproductive extranidal workers occurred is discussed. The best-supported hypothesis is that extranidal workers are old intranidal ones. Neither males nor inseminated workers were found in any smaples collected in the field or studied in the laboratory, which greatly strengthens earlier suggestions that Pristomyrmex pungens is the first-known ant to be obligately thelytokous. These findings indicate that Pristomyrmex pungens is no longer eusocial, although it has the highest form of social behavior of any thelytokous species; they also raise the question of whether or not there are factors promoting the loss of eusociality and sexuality in this species. Ecological factors are tentatively indicated, namely, the need to maintain large colonies in the face of a nomadic lifestyle involving frequent colony fragmentation.     

Functional morphology of the gonads of the articulate brachiopodTerebratulina retusa

The reproductive anatomy, and ultrastructural features of the gonads of the articulate brachiopodTerebratulina retusa (Linnaeus), are documented based on collections made between October 1985 and October 1986 from the Firth of Lorn, west coast of Scotland. This species is dioecious, and maturity is achieved in both sexes at shell lengths greater than ~5.5 mm. There is no obvious external sexual dimorphism except for slight differences in the coloration of the gonads; testes are white/cream, ovaries are yellow/orange. The gonads occur as four palmate lobes, a pair in each valve. Gonads are formed within a mantle sinus (vascula genitalia), which is an anterior extension of the coelom, that opens posteriorly into the visceral cavity and to the exterior via a pair of metanephridia. The latter serve as gonoducts during spawning. Gametes are borne on genital lamellae formed from a reticulate lattice of connective tissue. The lamellae are an extension of the ileoparietal band and are fused along one margin to the inner mantle membrane. Developing oocytes are closely affixed to the genital lamellae and originate from a pool of proliferating germ cells at its base. Vitellogenic oocytes that are at an advanced stage are released from the genital lamellae, but are retained within thevascula genitalia. Liberated oocytes continue to accumulate yolk and eventually occlude thevascula genitalia, before being spawned. Coelomocytes were identified within the gonads. In spent females these cells appear to be phagocytic and involved in the resorption of necrotic material, while in the male they may serve as nutritive cells.     

Bigger testes do work more: experimental evidence that testis size reflects testicular cell proliferation activity in the marine invertebrate,the free-living flatworm<Emphasis Type="Italic"> Macrostomum</Emphasis> sp.

Evolutionary theory predicts that more resources are allocated to sperm production when sperm competition is high. This prediction is supported by both comparative and experimental data on static measures of male allocation, such as testis and ejaculate size. However, resource allocation is a dynamic process, and it is therefore important to evaluate if the static measures reflect this. Such an evaluation has to our knowledge never been done. Immunocytochemical labelling with bromodeoxyuridine (BrdU) allows visualisation of cells in S-phase. BrdU becomes incorporated into cells if, and only if, they are actively undergoing DNA duplication, which is a dynamic process. The number of BrdU-positive cells in the testis can hence serve as a dynamic measure of male allocation, i.e. testicular activity. We evaluate the relationship between testis size and testicular activity in the marine flatworm Macrostomum sp. In a previous study, we showed that testis size is phenotypically plastic in this species, and that worms make larger testes when they are raised in larger groups. We use this plasticity to experimentally produce variation in testis size, and demonstrate that larger testes are associated with higher testicular activity. Moreover, testis size and testicular activity were related linearly. We have thus, for the first time, shown that testis size is a good measure of resource allocation to the male function. Moreover, increased testicular activity is probably one of the first steps in the upregulation of sperm production. It is thus expected that testicular activity is a more sensitive measure of short-term variation in male allocation than the commonly used static measures.Communicated by N. Wedell